Pyrazole derivative and use thereof for medical purposes

ABSTRACT

The present invention provides pyrazole derivatives, uses thereof for medical purposes and so on. More particularly, the present invention relates to pharmaceuticals useful for the prevention or treatment of constipation, which comprise as an active ingredient 3-(3-{4-[3-(β-D-glucopyranosyloxy)-5-isopropyl-1H-pyrazol-4-ylmethyl]-3-methylphenoxy}propylamino)-2,2-dimethylpropionamide, or a pharmaceutically acceptable salt thereof. The pharmaceuticals of the present invention exert an effect of increasing the frequency of bowel movement or the like, and are useful for the prevention or treatment of constipation.

This application is a Continuation of U.S. application Ser. No.14/399,093, filed on Nov. 5, 2014, which is a National Stage ofInternational Application No. PCT/JP2013/062755, filed on May 2, 2013,which claims priority to Japanese priority application No. 2012-105847filed on May 7, 2012, which are hereby incorporated by reference.

TECHNICAL FIELD

The present invention relates to pharmaceuticals which are useful forthe prevention or treatment of constipation.

More particularly, the present invention relates to pharmaceuticalswhich are useful for the prevention or treatment of constipation andwhich comprise as an active ingredient a compound (chemical name:3-(3-{4-[3-(β-D-glucopyranosyloxy)-5-isopropyl-1H-pyrazol-4-ylmethyl]-3-methylphenoxy}propylamino)-2,2-dimethylpropionamide,hereinafter sometimes referred to as “Compound 1”) represented by theformula:

or a pharmaceutically acceptable salt thereof

BACKGROUND ART

Normally, bowel movement occurs habitually and dose not prevent someonefrom leading daily life. However, comfortable bowel movement ofsufficient volume becomes difficult for some reasons and a conditionassociated with a physical suffering is caused. It is calledconstipation. Constipation is generally associated with straining duringbowel movement, hard feces, decreased frequency of bowel movement,sensation of incomplete evacuation, bloating, prolonged transit time offood/stool in the entire gastrointestinal tract or colon and so on.

Constipation is classified into acute constipation and chronicconstipation based on a period of medical history. In addition,constipation is classified into organic constipation and functionalconstipation based on its etiology. Organic constipation is a conditionthat it is difficult to defecate due to organic disorders such asstricture, obstruction and so on of gastrointestinal tract caused bycolonic cancer, colonic polyps, uterine fibroid and so on. Furthermore,functional constipation is further classified into symptomaticconstipation, drug-induced constipation and other constipation.Symptomatic constipation is constipation secondary to diseases otherthan gastrointestinal diseases. Drug-induced constipation isconstipation which is secondarily caused by drugs, and it is known thatit is caused by the administration of drugs which have an antimotilityeffect, such as opioid, anticholinergic drugs and the like. Functionalconstipation which is not symptomatic constipation or drug-inducedconstipation is the most common type, it is also called chronicidiopathic constipation (CIC), and it is caused by various reasons suchas changes in eating habits and living environment, psychologicalfactors and so on. Chronic constipation can be also classified into slowtransit constipation and outlet obstruction based on reasons ofconstipation. Slow transit constipation is a condition in which thepassage of the stool through the proximal colon to distal colon isimpaired due to the decrease of colonic smooth muscle contraction,diminished peristalsis and the like, and outlet obstruction is acondition in which it is not possible to defecate due to impairedfunction of defecation even though the stool is transferred to rectum.Irritable bowel syndrome with constipation (IBS-C) is constipation inwhich gastrointestinal symptom dominated by abdominal pain, abdominaldiscomfort and stool abnormality is continued without organic changes inthe gastrointestinal tract, and some patients of functional constipationmay be classified as IBS-C (see Non-patent literatures 1-3).

The treatment of constipation includes life therapy, drug therapy,behavior therapy and surgical therapy. As the first choice in thetreatment, life therapy involving a correction of irregular dietaryhabits, correction of bowel habits, and sufficient intake of high-fiberfood and water is a basic treatment. Drug therapy is indicated inpatients whose symptoms of constipation do not improve with lifetherapy.

In principle, drug therapy is initiated with drugs having a mild effectsuch as osmotic laxatives and bulk-forming laxatives which increase avolume of gut content. Irritant laxatives, prokinetic agents and thelike are used when the effect of the above drugs is insufficient.Osmotic laxatives and bulk-forming laxatives are less addictive and canbe administered for a long time. However, it is important to care forrenal disorder, abnormal electrolyte level in the blood, hypermagnesemiaof renal disorder or the like. In addition, it is known that addictionsand inflammatory changes on intestinal mucosa may be caused by continuedadministration of irritant laxatives.

Lubiprostone is known as a new therapeutic agent for constipation (seeNon-patent literature 4). Lubiprostone is sold as a therapeutic agentfor chronic constipation (except for constipation due to organicdiseases) in Japan. Lubiprostone is also sold as a therapeutic agent forchronic idiopathic constipation and irritable bowel syndrome withconstipation, and is approved as a therapeutic agent for opioid-inducedconstipation (OIC) in the U.S.A.

Therefore, it is said that the drugs for the prevention or treatment ofconstipation are not sufficient now, and a drug for the prevention ortreatment of constipation having a new mechanism of action, which causesfewer adverse reactions, has been strongly desired.

Compound 1 or a pharmaceutically acceptable salt thereof is known to beuseful as an agent for the prevention or treatment of a diseaseassociated with hyperglycemia such as diabetes, abnormal glucosetolerance, impaired fasting glucose, diabetic complication, obesity andso on (see Patent literatures 1-3).

However, it is not known that Compound 1 or a pharmaceuticallyacceptable salt thereof is useful as an agent for the prevention ortreatment of constipation.

-   Patent literature 1: International publication No. WO2004/018491-   Patent literature 2: International publication No. WO2009/084531-   Patent literature 3: International publication No. WO2009/128421-   Non-patent literature 1: Tetsuji Kitahora and 4 persons, Bessatsu    Nippon Rinsho Shinryouikibetsusyoukougun series, 2009, Vol. 12, p.    422-427-   Non-patent literature 2: Kouji Komori and 4 persons, Bessatsu Nippon    Rinsho Shinryouikibetsusyoukougun series, 2009, Vol. 12, p. 433-435-   Non-patent literature 3: George F. Longstreth et al,    Gastroenterology, 2006, Vol. 130, p. 1480-1491-   Non-patent literature 4: S. Fukudo et al, Neurogastroenterology and    Motility, 2011, Vol. 23, p. 544-e205

DISCLOSURE OF THE INVENTION Problems to be Solved by the Invention

A problem of the present invention is to provide pharmaceuticals and thelike which are useful for the prevention or treatment of constipation.

Means for Solving the Problems

The present invention relates to pharmaceuticals for use in theprevention or treatment of constipation, which comprise as an activeingredient Compound 1, or a pharmaceutically acceptable salt thereof.

That is, the present invention relates to:

[1] a pharmaceutical for use in the prevention or treatment ofconstipation, which comprises as an active ingredient3-(3-{4-[3-(β-D-glucopyranosyloxy)-5-isopropyl-1H-pyrazol-4-ylmethyl]-3-methylphenoxy}propylamino)-2,2-dimethylpropionamide,or a pharmaceutically acceptable salt thereof;

[2] the pharmaceutical as described in the above [1], which comprises asan active ingredientbis[3-(3-{4-[3-(β-D-glucopyranosyloxy)-5-isopropyl-1H-pyrazol-4-ylmethyl]-3-methylphenoxy}propylamino)-2,2-dimethylpropionamide]monosebacate;

[3] the pharmaceutical as described in the above [1] or [2], wherein theconstipation is functional constipation;

[4] the pharmaceutical as described in the above [3], wherein thefunctional constipation is chronic idiopathic constipation;

[5] the pharmaceutical as described in the above [3], wherein thefunctional constipation is drug-induced constipation; and the like.

Effect of the Invention

The pharmaceuticals of the present invention exert an effect ofincreasing the frequency of bowel movement or the like, and are usefulfor the prevention or treatment of constipation.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 shows the result of Example 1, which includes the frequency ofbowel movement within 24 hours after the administration. In the FIGURE,each bar chart shows a value of Normal group (Normal), Control group(Control), the group administered with 1 mg/kg of Compound 2, the groupadministered with 3 mg/kg of Compound 2, the group administered with 10mg/kg of Compound 2, the group administered with 0.1 mg/kg oflubiprostone, or the group administered with 0.7 mg/kg of lubiprostonefrom the left respectively. The vertical axes show the frequency ofbowel movement (which is the frequency indicating bowel movement in theobservation three times a day) (The data indicate the mean±standarderror of 10 examples per group.). * shows a significant difference withthe Control group.

MODE FOR CARRYING OUT THE INVENTION

As the pharmaceutically acceptable salt of Compound 1, an acid additivesalt with a mineral acid such as hydrochloric acid, hydrobromic acid,nitric acid, phosphoric acid and the like, an acid additive salt with anorganic acid such as formic acid, acetic acid, methanesulfonic acid,benzenesulfonic acid, p-toluenesulfonic acid, propionic acid, citricacid, succinic acid, tartaric acid, fumaric acid, butyric acid, oxalicacid, malonic acid, maleic acid, lactic acid, malic acid, carbonic acid,glutamic acid, aspartic acid, benzoic acid, sebacic acid, pamoic acidand the like, and so on can be illustrated. More preferably,monosebacate of Compound 1 (chemical name:bis[3-(3-{4-[3-(β-D-glucopyranosyloxy)-5-isopropyl-1H-pyrazol-4-ylmethyl]-3-methylphenoxy}propylamino)-2,2-dimethylpropionamide]monosebacate;hereinafter sometimes referred to as “Compound 2”), hemifumaratedihydrate of Compound 1 (chemical name:3-(3-{4-[3-(β-D-glucopyranosyloxy)-5-isopropyl-1H-pyrazol-4-ylmethyl]-3-methylphenoxy}propylamino)-2,2-dimethylpropionamidehemifumarate dihydrate; hereinafter sometimes referred to as “Compound3”) and the like can be illustrated.

Compound 1 or a pharmaceutically acceptable salt thereof of the presentinvention also includes a solvate thereof with a pharmaceuticallyacceptable solvent (such as water, ethanol or the like).

Compound 1 or a pharmaceutically acceptable salt thereof of the presentinvention can be also prepared by a method described in patentliteratures 1-3 or a similar method thereto.

Compound 1 of the present invention may be converted to a prodrugappropriately and be used. For example, a prodrug of Compound 1 can beprepared by introducing an appropriate group forming a prodrug into anyone or more groups selected from a hydroxy group, an amino group and anamino group of the pyrazole ring of Compound 1 using a correspondingreagent to produce a prodrug such as a halide compound or the like inthe usual way, and then by suitably isolating and purifying in the usualway as occasion demands. As a group forming a prodrug, for example, agroup as described in “Development of medicine” 1990, Vol. 7, p.163-198, published by Hirokawa shoten can be illustrated.

The pharmaceuticals of the present invention can be administered asvarious formulations which include oral forms, for example, such astablets, capsules, granules, powders, fine granules, dry syrups and thelike, and parenteral forms such as liquid forms, ointment forms,suppositories and the like.

The pharmaceuticals of the present invention can be prepared as variousformulations by admixing or diluting/dissolving an active ingredientwith an appropriate pharmaceutical carrier such us excipients,disintegrators, binders, lubricants, diluents, buffers, tonicity agents,preservatives, wetting agents, emulsifiers, dispersants, stabilizers,solubilization agents and the like using conventional methods.

The pharmaceuticals of the present invention can be also administered incombination with another drug which is used in the treatment ofconstipation. As another drug, for example, bulk-forming laxatives suchas carmellose sodium and the like, osmotic laxatives such as magnesiumoxide and the like, irritant laxatives such as sodium picosulfatehydrate and the like, enemas such as glycerin and the like,suppositories such as sodium hydrogen carbonate/anhydrous sodiumdihydrogen phosphate and the like, tuning agents of gastrointestinalmotility such as trimebutine maleate and the like, chloride channelactivators such as lubiprostone and the like, guanylyl cyclase receptoragonists such as linaclotide and the like, μ-opioid receptor antagonistssuch as methylnaltrexone and the like, bile acid transporter inhibitorssuch as elobixibat and the like, serotonin 4 receptor agonists such asprucalopride and the like, and so on can be illustrated.

When the pharmaceuticals of the present invention are used incombination with the above drugs, the present invention includes allsimultaneous administration as a single formulation, simultaneousadministration as separate formulations by the same administrationpathway or different pathways, and administration at different times asseparate formulations by the same administration pathway or differentpathways.

The dosage of an active ingredient of the present invention isappropriately decided depending on the body weight, age, sex, degree ofdisorders of each patient and the like. The dosage in an adult human canbe decided within the range of, for example, 0.1 to 160 mg per day, 1 to60 mg per day, 2 to 60 mg per day, 2 to 40 mg per day, 2 to 20 mg perday or 2 to 10 mg per day in the case of oral administration, and thedaily dose can be divided into one, two or three times per day andadministered.

In addition, for example, 1 mg once daily, 1 mg twice daily, 1 mg threetimes daily, 2 mg once daily, 2 mg twice daily, 2 mg three times daily,2.5 mg once daily, 2.5 mg twice daily, 2.5 mg three times daily, 5 mgonce daily, 5 mg twice daily, 5 mg three times daily, 10 mg once daily,10 mg twice daily, 10 mg three times daily, 15 mg once daily, 15 mgtwice daily, 15 mg three times daily, 20 mg once daily, 20 mg twicedaily, 20 mg three times daily, 40 mg once daily, 40 mg twice daily, 80mg once daily, or 80 mg twice daily can be administered.

Furthermore, the first dosage is selected from, for example, 1 mg, 2 mg,2.5 mg, 5 mg, 10 mg or 20 mg, and then the dosage can be increased ordecreased gradually depending on the sensitivity, the degree ofdisorders or the like of each patient.

The dosage can be decided within the range of, for example, 0.05 to 80mg per day in the case of parenteral administration.

The pharmaceuticals of the present invention can be also administeredbefore a meal, after a meal or with a meal, preferably administeredafter a meal.

In the present invention, functional constipation is constipation otherthan organic constipation among constipation. Chronic idiopathicconstipation (CIC) is constipation other than symptomatic constipationand drug-induced constipation among functional constipation, andincludes atonic constipation, spastic constipation, rectal constipationand the like.

Also, in the present invention, chronic constipation includes chronicorganic constipation and chronic functional constipation. Therefore,chronic constipation includes chronic idiopathic constipation (CIC),symptomatic constipation, drug-induced constipation, irritable bowelsyndrome with constipation (IBS-C) and chronic organic constipation.

Symptomatic constipation is constipation secondary to diseases otherthan gastrointestinal diseases among functional constipation, andincludes constipation which is caused by endocrine diseases such ashypothyroidism, pheochromocytoma, hypopituitarism, hyperparathyroidismand so on, metabolic diseases such as amyloidosis, uremia and so on,poisonings such as lead poisoning, arsenic poisoning and so on,neurologic diseases such as Parkinson's disease, cerebrovasculardisorder, brain tumor, multiple sclerosis and so on, connective tissuediseases such as scleroderma and so on, and anus diseases such asperianal abscess and so on, and the like.

Drug-induced constipation is constipation which is secondarily caused bydrugs among functional constipation, and includes constipation which iscaused by the administration of a drug having an antimotility effectsuch as opioid and so on, an anticholinergic drug and the like (forexample opioid-induced constipation).

Irritable bowel syndrome with constipation (IBS-C) is constipation inwhich gastrointestinal symptom dominated by abdominal pain, abdominaldiscomfort and stool abnormality is continued without organic changes inthe gastrointestinal tract, and is included in the above functionalconstipation.

The diagnosis for functional constipation can be also measured forexample by Rome III Diagnostic Criteria (see Non-patent literature 3, inspecific p. 1486).

The diagnosis for IBS-C can be also measured for example by said RomeIII Diagnostic Criteria (see Non-patent literature 3, in specific p.1481-1482).

The pharmaceuticals of the present invention can improve one or two ormore of the symptoms of constipation (frequency of bowel movements,sensation of incomplete evacuation, straining, stool form, abdominalbloating, abdominal discomfort and so on).

EXAMPLE

The present invention is further illustrated in more detail by way ofthe following Examples. However, the present invention is not limitedthereto.

Example 1 Improvement Effect in Constipation Model 1 1. PreparationProcedure of Dosing Solution (1) Preparation Procedure of Test Compounds

Compound 2 was weighed and was dissolved in distilled water atpreparation concentrations of 0.5, 1.5 and 5 mg/mL to prepare testcompounds.

(2) Preparation Procedure of Control Substances

Lubiprostone (TLC Pharma Chem) was weighed and was suspended in 0.5%methylcellulose at preparation concentrations of 0.05 and 0.35 mg/mL toprepare control substances.

(3) Preparation Procedure of Reagent for Preparing the Model

Loperamide hydrochloride (Wako pure chemical) was weighed at 0.3, 0.5,1.0, 2.0, 4.0 or 8.0 mg/kg for each animal and was filled into a gelatincapsule.

(4) Preparation Procedure of a Solution of Mixed Carbohydrate

A soluble starch, sucrose and lactose monohydrate were weighed at a rateof 6:3:1 and were dissolved in distilled water which was approximately80% of the amounts of preparation on heating, and then the mixture wasadded with distilled water to prepare 0.4 g/mL solution of mixedcarbohydrate.

2. Method (1) Loperamide-Induced Dog Constipation Model

Loperamide hydrochloride which was filled into a gelatin capsule wasorally administered to dogs (beagle, male, 13-14 months old, 10 dogs,Kitayama-labesu). The dosage was increased from 0.3 mg/kg, and was setat 2.0, 4.0 or 8.0 mg/kg based on the bowel movement condition of eachdog. Dogs whose wet fecal weights during 24 hours after theadministration of loperamide hydrochloride were significantly lower thanthe wet fecal weight during 24 hours of the non-treated dogs (Normalgroup) (fecal weight after administration<(average fecal weight withoutadministration−2× standard deviation)) were subjected to the tests asconstipation model animals.

(2) Experimental Procedure

Tests were performed as a full cross-over trial. More than 5-day washoutperiod was provided between each test, and a recovery of stool form wasidentified, and then the next test was performed.

A gelatin capsule which was filled with loperamide hydrochloride wasadministered orally at around 9 a.m. on the first and second days ineach test.

Compound 2 (1 mg/kg, 3 mg/kg or 10 mg/kg), lubiprostone (0.1 mg/kg or0.7 mg/kg) or distilled water (2 mL/kg) was administered orally ataround 4 p.m. on the second day using a glass syringe and an oralcatheter, and then 0.4 g/mL solution of mixed carbohydrate wasadministered orally at 50 mL/body. The feces were observed at 17 hours,21 hours and 24 hours after the administration, and the feces within 24hours after the administration were collected and weighed, and it wasset as the wet fecal weight. With regard to the frequency of bowelmovement, it was counted as one bowel movement when the feces wereobserved at each observation point regardless of the size of the fecalvolume. The stool form of feces was scored in seven categories based onBristol Stool Form Scale. The collected feces were dried fully, and thenthe feces were weighed, and it was set as the dry fecal weight.

(3) Data Processing

The statistical analysis was made by testing the uniformity of varianceusing Bartlett's method, and multiple comparison between the group whichwas given distilled water (Control group) and each administered groupwas made using Dunnett's method when variance was uniform, and multiplecomparison was made using Steel's method when variance was not uniform.In each case, significance level<5% was considered as significantdifference.

3. Result

The data obtained from the tests were evaluated statistically. The meansof wet fecal weights, dry fecal weights and frequencies of bowelmovements within 24 hours after the administration of 10 subjects ineach group are shown in Tables 1-3 and FIG. 1. Both Compound 2 andlubiprostone dose-dependently increased the wet fecal weight (Table 1),dry weight (Table 2) and frequency of bowel movements (Table 3 and FIG.1), and these effects in the group administered with 3 mg/kg or more ofCompound 2 and 0.1 mg/kg or more of lubiprostone were significantlyhigher than in Control group. In this test, increases of loose stool anddiarrhea were not observed.

It was suggested that Compound 2 had the effects of increasing the fecalvolume and the frequency of bowel movement in constipation model, andwas useful as an agent for the prevention or treatment of constipationfrom the result of Example 1.

TABLE 1 Wet fecal weight within 24 hours after the administration GroupWet fecal weight (g) Normal group (non-treated) 136.2 Control group(distilled water) 2.8 The group administered with 1 mg/kg 29.5 ofCompound 2 The group administered with 3 mg/kg 60.9 of Compound 2 Thegroup administered with 10 mg/kg 77.2 of Compound 2 The groupadministered with 0.1 mg/kg 18.7 of lubiprostone The group administeredwith 0.7 mg/kg 55.4 of lubiprostone

TABLE 2 Dry fecal weight within 24 hours after the administration GroupDry fecal weight (g) Normal group (non-treated) 47.1 Control group(distilled water) 1.0 The group administered with 1 mg/kg 10.4 ofCompound 2 The group administered with 3 mg/kg 21.6 of Compound 2 Thegroup administered with 10 mg/kg 27.3 of Compound 2 The groupadministered with 0.1 mg/kg 8.3 of lubiprostone The group administeredwith 0.7 mg/kg 21.5 of lubiprostone

TABLE 3 Frequency of bowel movement within 24 hours after theadministration Frequency of bowel movement Group (time) Normal group(non-treated) 2.1 Control group (distilled water) 0.2 The groupadministered with 1 mg/kg of 0.5 Compound 2 The group administered with3 mg/kg of 1.2 Compound 2 The group administered with 10 mg/kg of 1.5Compound 2 The group administered with 0.1 mg/kg of 0.9 lubiprostone Thegroup administered with 0.7 mg/kg of 1.0 lubiprostone

Example 2 Improvement Effect in Constipation Model 2 1. PreparationProcedure of Dosing Solution (1) Preparation Procedure of Test Compounds

Compound 2 was weighed and was dissolved in distilled water atpreparation concentrations of 3, 10 and 30 mg/mL as the free drug toprepare test compounds.

(2) Preparation Procedure of Control Substances

0.5% methylcellulose (0.5% MC) was used as a vehicle for preparation oflubiprostone. Lubiprostone was weighed and was suspended in 0.5% MC atpreparation concentrations of 1, 3 and 10 mg/mL to prepare controlsubstances.

(3) Preparation Procedure of a Positive Control Substance

Magnesium sulfate (MgSO₄) (Wako pure chemical) was weighed and wasdissolved in distilled water at a preparation concentrations of 200mg/mL to prepare a positive control substance.

(4) Preparation Procedure of a Solution of Mixed Carbohydrate

A soluble starch, sucrose and lactose monohydrate were weighed at a rateof 6:3:1 and were dissolved in distilled water which was approximately80% of the amounts of preparation on heating, and then the mixture wasadded with distilled water to prepare 0.4 g/mL solution of mixedcarbohydrate.

2. Low-Fiber Diet

A low-fiber diet was prepared based on the literature (Kakino et al. BMCComplementary and Alternative Medicine 2010, 10: 68).

3. Method (1) Preparation of a Rat Model of Low-Fiber Diet-InducedConstipation.

Rats were fed on the low-fiber diet for 1-2 weeks to induceconstipation. Rats whose fecal weights during 24 hours weresignificantly lower than the fecal weight during 24 hours of the normalfeed group (Normal group) (fecal volume when fed on the low-fiberdiet<(average fecal volume of normal feed group−2× standard deviation))were determined as constipation condition, and the rats which were fedon the low-fiber diet for 1 more week were subjected to the tests as amodel of chronic constipation.

(3) Experimental Procedure

Compound 2 (3 mg/kg, 10 mg/kg or 30 mg/kg as free drug), lubiprostone (1mg/kg, 3 mg/kg or 10 mg/kg), MgSO₄ (2000 mg/kg) or a vehicle (distilledwater: Control group 1, 0.5% MC: Control group 2) was administeredorally at around 9 a.m. using a 1 mL glass syringe and a stomach tube.Then, 0.4 g/mL solution of mixed carbohydrate was administered orally at2 mL/body to Control group 1 and the groups administered with Compound2. Distilled water was administered to the normal feed group. Testgroups and the numbers of examples are shown in Table 4.

The feces were observed at 4, 8, 12 and 24 hours after theadministration, and the feces were collected and weighed, and it was setas the fecal weight. The sum of the fecal weights at the collectiontimes was set as the fecal weight at 24 hours after the administration.

TABLE 4 Test group and number of animal Test Test group Number of animal(i) Normal feed group 4 (ii) Control group 1 4 The group administeredwith 3 mg/kg 4 of Compound 2 The group administered with 10 mg/kg 4 ofCompound 2 The group administered with 30 mg/kg 4 of Compound 2 Thegroup administered with MgSO₄ 4 (iii) Control group 2 5 The groupadministered with 1 mg/kg 5 of lubiprostone The group administered with3 mg/kg 5 of lubiprostone The group administered with 10 mg/kg 5 oflubiprostone

(4) Data Processing

The statistical analysis was made by testing the uniformity of varianceusing Bartlett's method, and multiple comparison between Control group 1and the group administered with Compound 2 or Control group 2 and thegroup administered with lubiprostone was made using Dunnett's methodwhen variance was uniform respectively, and multiple comparison was madeusing Steel's method when variance was not uniform. In each case,significance level<5% was considered as significant difference.

4. Result

The combined and summarized data obtained from the normal feed group(i), the tests of Compound 2 and magnesium sulfate administration (ii)and the tests of lubiprostone administration (iii) are shown in Table 5.

Both Compound 2 and lubiprostone dose-dependently increased the fecalweight during 24 hours after the administration, and these effects inthe groups administered with 10 mg/kg or more of Compound 2 and 10 mg/kgof lubiprostone were significantly higher than in each Control group.The increase of the fecal weight was also shown in the groupadministered with MgSO₄ which is a positive control. Moreover, waterystool was reported in the groups administered with 10 mg/kg or more ofCompound 2, 3 mg/kg or more of lubiprostone and MgSO₄.

It was suggested that Compound 2 had the effects of increasing the fecalvolume in constipation model, and was useful as an agent for theprevention or treatment of constipation from the result of Example 2.

TABLE 5 Fecal weight within 24 hours after the administration Test groupFecal weight (g) Normal feed group 8.33 Control group 1 0.53 The groupadministered with 3 mg/kg 1.20 of Compound 2 The group administered with10 mg/kg 4.20 of Compound 2 The group administered with 30 mg/kg 4.93 ofCompound 2 The group administered with MgSO₄ 3.70 Control group 2 0.58The group administered with 1 mg/kg 0.60 of lubiprostone The groupadministered with 3 mg/kg 0.95 of lubiprostone The group administeredwith 10 mg/kg 1.25 of lubiprostone

Example 3 Single-Dose Study in Healthy Adult Males 1. Method

Compound 2 at a single dose of 2, 5, 10, 20, 40, 80 or 160 mg (free drugequivalent), or placebo was orally administered just immediately beforebreakfast to healthy adult males. For the administration of Compound 2,tablets containing 1, 5 or 10 mg in free drug equivalent were used. Thetest period was set between the administration and the discharge 48hours after the administration. Frequencies of bowel movements werechecked during consultation by a doctor, and the adverse events(gastrointestinal disorders) were defined when an abnormal finding(diarrhea or loose stool) was seen based on the stool form and thefinding.

2. Result

In the groups administered with Compound 2, increases in the frequenciesof bowel movement were observed (Table 6), and stool forms weredetermined at Bristol Stool Form Scale type 6 or 7 for many subjects inthe 80 mg and 160 mg groups. Therefore, it was shown that Compound 2made the stool soft and increased the frequency of bowel movement alsoin humans. The number of subjects with abdominal bloating, abdominalpain and diarrhea recoded as gastrointestinal disorders are shown inTable 7, but the severity was mild in all the cases.

TABLE 6 Frequency of bowel movement after the administration Compound 2(mg) Dose 2 5 10 20 40 80 160 Placebo Number of 6 6 6 6 6 6 6 14subjects (subjects) Mean 1.7 1.0 2.2 2.2 1.8 4.8 5.2 1.7 (time) SD 0.80.9 1.2 0.8 1.7 3.5 1.2 1.1

TABLE 7 Number of subject with gastrointestinal disorders Compound 2(mg) Dose 2 5 10 20 40 80 160 Placebo Number of 6 6 6 6 6 6 6 14subjects Abdominal 0 0 0 0 0 2 3 0 bloating Abdominal 0 0 0 0 0 4 3 0pain Diarrhea 1 0 2 2 2 5 6 0

Example 4 Multiple-Dose Study in Healthy Adult Males 1. Method

Compound 2 at a dose of 2, 5, 10 or 20 mg (free drug equivalent), aplacebo, or miglitol at a dose of 50 mg was repetitively administeredorally once daily just immediately before breakfast on administrationDays 1 and 13, and 3 times daily just immediately before every meal onadministration Days 3 to 12 to healthy adult males. For theadministration of Compound 2, tablets containing 1, 5 or 10 mg in freedrug equivalent were used. The test period was set betweenadministration Day 1 and Day 15. Frequencies of bowel movements werechecked during consultation by a doctor, and the adverse events(gastrointestinal disorders) were defined when an abnormal finding(diarrhea or loose stool) was seen based on the stool form and thefinding. In this regard, the frequencies of bowel movements in Table 8show the daily means during fifteen days (time/day).

2. Result

There was a tendency of dose-dependent increase in the number of thesubjects who had bowel movements with Bristol Stool Form Scale type 6 ortype 7 and in the frequency thereof. The numbers of the subjects withdiarrhea and abdominal pain recoded as gastrointestinal disorders areshown in Table 9. However, the severity of each subject was mild and thedisorders disappeared or recovered without treatment.

TABLE 8 Daily frequency of bowel movement after the administrationCompound 2 (mg) Dose 2 5 10 20 Placebo Miglitol Number of 8 8 8 8 8 8subjects (subjects) Mean (time/day) 1.4 1.1 2.0 1.7 1.0 1.0 SD 0.6 0.51.2 1.0 0.4 0.4

TABLE 9 Number of subject with gastrointestinal disorders Compound 2(mg) Dose 2 5 10 20 Placebo Miglitol Number of 8 8 8 8 8 8 subjectsDiarrhea 6 5 5 6 3 2 Abdominal pain 0 0 0 1 0 0

Example 5 Clinical Trial in Patients with Chronic Constipation 1. Method

After observation for 2 weeks, Compound 2 or a placebo was administeredorally after eating to 79 patients with chronic constipation for 4weeks. Patients who had steady symptoms for long periods were selectedas constipation patients using diagnostic criteria of functionalconstipation based on Rome III as a reference.

Each administered group is as follows:

2 mg TID group: administration of 2 mg of Compound 2, 3 times daily

-   -   (after breakfast, after lunch, after dinner)

20 mg QD group: administration of 20 mg of Compound 2, 1 time daily

-   -   (after breakfast)

20 mg BID group: administration of 20 mg of Compound 2, 2 times daily

-   -   (after breakfast, after dinner)

20 mg TID group: administration of 20 mg of Compound 2, 3 times daily

-   -   (after breakfast, after lunch, after dinner)

Placebo group: administration of placebo, 3 times daily

-   -   (after breakfast, after lunch, after dinner)

For the administration of Compound 2, tablets containing 1 or 10 mg infree drug equivalent of Compound 2 were used.

If the subjects complained that further administration of 2 tablets eachtime was difficult due to the increase in the frequency of spontaneousbowel movements or softened stool form causing problems in daily life,the dose was switched to 1 tablet each time in the week 1 visit or week2 visit by the doctor and the administration was continued.

2. Evaluation Items

Frequency of spontaneous bowel movements, frequency of complete bowelmovements (frequency of spontaneous bowel movements without sensation ofincomplete evacuation), percentage of patients who had a bowel movementwithin 24 hours after the first administration, percentage of patientswho had a bowel movement within 48 hours after the first administration,time until the first spontaneous bowel movement, stool form (BristolStool Form Scale) and so on were evaluated.

3. Result (1) Subject 1

It was shown that the frequencies of spontaneous bowel movements perweek of the subject 1 who was given 2 mg of Compound 2, 3 times daily (2mg TID group) were 1.1, 7.0, 7.0, 11.0 and 10.5 (time/week) during theobservation period, administration week 1, week 2, week 3 and week 4,respectively.

In addition, it was shown that the Bristol Stool Form Scales of thesubject 1 were 1.5, 3.7, 3.7, 4.1 and 3.8 (mean/week) during theobservation period, administration week 1, week 2, week 3 and week 4,respectively.

Also, the time until the first bowel movement was 5 hours and 20minutes.

(2) Subject 2

It was shown that the frequencies of spontaneous bowel movements perweek of the subject 2 who was given 20 mg of Compound 2, 1 time daily(20 mg QD group) were 1.8, 12.0, 13.0 and 8.0 (time/week) during theobservation period, administration week 1, week 2 and week 3,respectively.

In addition, it was shown that the Bristol Stool Form Scales of thesubject 2 were 3.0, 4.6, 4.0 and 3.4 (mean/week) during the observationperiod, administration week 1, week 2 and week 3, respectively.

Also, the the time until the first bowel movement was 24 hours and 20minutes.

(3) Subject 3

It was shown that the frequencies of spontaneous bowel movements perweek of the subject 3 who was given 20 mg of Compound 2, 2 times daily(20 mg BID group) and whose dose was then switched to 10 mg of Compound2, 2 times daily at the day after the 1-week administration were 2.2,21.0, 6.0, 6.0 and 5.0 (time/week) during the observation period,administration week 1, week 2, week 3 and week 4, respectively.

In addition, it was shown that the Bristol Stool Form Scales of thesubject 3 were 2.0, 5.9, 4.0, 4.3 and 4.2 (mean/week) during theobservation period, administration week 1, week 2, week 3 and week 4,respectively.

Also, the the time until the first bowel movement was 1 hour and 20minutes.

It was shown that Compound 2 facilitated the spontaneous bowel movementsand had an improvement effect on the stool form in patients with chronicconstipation, and was useful as an agent for the treatment of chronicconstipation from the result of Example 5. In this regard, no subjectswith hypoglycemia were reported in the groups administered with Compound2.

INDUSTRIAL APPLICABILITY

The pharmaceuticals of the present invention are very useful for theprevention or treatment of constipation.

1. A method for the treatment of constipation, comprising administeringto a patient in need thereof a pharmaceutical composition comprising asan active ingredient3-(3-(3-{4-[3-(β-D-glucopyranosyloxy)-5-isopropyl-1H-pyrazol-4-ylmethyl]-3-methylphenoxy}propylamino)-2,2-dimethylpropionamide,or a pharmaceutically acceptable salt thereof, and an appropriatepharmaceutical carrier.
 2. The method for the treatment of constipationas claimed in claim 1, wherein the active ingredient isbis[3-(3-{4-[3-(β-D-glucopyranosyloxy)-5-isopropyl-1H-pyrazol-4-ylmethyl]-3-methylphenoxy}propylamino)-2,2-dimethylpropionamide]monosebacate.3. The method for the treatment of constipation as claimed in claim 1,wherein the constipation is functional constipation.
 4. The method forthe treatment of constipation as claimed in claim 3, wherein thefunctional constipation is chronic idiopathic constipation.
 5. Themethod for the treatment of constipation as claimed in claim 3, whereinthe functional constipation is drug-induced constipation.
 6. The methodfor the treatment of constipation as claimed in claim 2, wherein theconstipation is functional constipation.